Entirely Carbohydrate Vaccine Constructs and Their Application in Probing Glycoim

全碳水化合物疫苗构建体及其在检测糖蛋白中的应用

• 批准号: 8730098
• 负责人: Peter R Andreana
• 金额: $ 29.29万
• 依托单位: UNIVERSITY OF TOLEDO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8730098
• 关键词: Affinity; Anaerobic Bacteria; Antibodies; Antibody Specificity; Antigens; Area; B-Lymphocytes; Bacterial Polysaccharides; Binding; Biological Assay; Breast Cancer Cell; Breast Carcinoma; C57BL/6 Mouse; CD4 Positive T Lymphocytes; Cancer Vaccines; Cancer cell line; Carbohydrate Chemistry; Carbohydrates; Carrier Proteins; Cell surface; Cells; Cellular Immunity; Charge; Chemicals; Chemistry; Complement; Complement-Dependent Cytotoxicity; Complex; Conjugate Vaccines; Development; Enzyme-Linked Immunosorbent Assay; Evaluation; Event; Flow Cytometry; Glycoproteins; Goals; Histocompatibility Antigens Class II; Human; Humoral Immunities; Hydrazones; Immune; Immune response; Immune system; Immunity; Immunization; Immunoglobulin G; Immunoglobulin M; In Vitro; Knowledge; Lactate Dehydrogenase; Lead; Lipids; MCF7 cell; Malignant Neoplasms; Mediating; Medicine; Methodology; Methods; Modeling; Modification; Monosaccharides; Mus; Neoplasm Metastasis; Oximes; Pathway interactions; Peptides; Polysaccharides; Prevention therapy; Preventive; Process; Property; Proteins; Research; Role; Route; Scheme; Serum; Site; Solid Neoplasm; Specificity; Structure; Subcutaneous Injections; Survival Rate; T-Lymphocyte; Testing; Therapeutic; Tumor Cell Line; Tumor-Associated Carbohydrate Antigens; U251; Vaccine Research; Vaccines; Work; arm; base; cancer cell; cancer therapy; cell killing; cytotoxicity; design; disorder prevention; efficacy testing; immune function; immunogenic; immunogenicity; improved; in vivo; innovation; insight; killings; novel; oxidation; programs; research study; response; sugar; tool; tumor; tumor growth; vaccine candidate; vaccine development

DESCRIPTION (provided by applicant): The principal goal of this research program is to develop practical synthetic approaches for the modification of a unique capsular zwitterionic polysaccharide (ZPS), PS A1, and conjugate tumor-associated carbohydrate antigens (TACAs) to develop entirely carbohydrate vaccine constructs. PS A1 is a structurally well- characterized polysaccharide containing a repeating tetrasaccharide core isolated from the capsular polysaccharide complex (CPC) of the anaerobe Bacterioides fragilis NCTC 9343. It was determined that PS A1 could coax an immune system to elicit cellular and humoral immune functions via T- and B-cell involvement that influenced helper and memorial response events - a paradigm shifting discovery. What is not entirely clear is the immunogenicity of entirely carbohydrate-based constructs as an alternative approach to glycoprotein conjugates for tumor vaccine development. This research program, therefore, aims to study and assess the innovative vaccine constructs using three specific aims: 1) To develop efficient conjugation methods for the construction of structurally well-defined TACA-PS A1 conjugate vaccines, 2) To study the immunological properties of TACA-PS A1 conjugates in mice and evaluate the specificity and cytotoxicity of TACA-PS A1- raised Abs to human cancer cell lines, and 3) To evaluate, in vivo, the efficacy of TACA-PS A1-invoked immunity to treat tumors. Anti-sera, obtained from murine immunizations will be used to determine total antibody (Ab) titers and Ab isotypes by ELISA, relaying information about immunity. Furthermore, ELISA will be used to determine Ab specificity against conjugated TACAs. Flow cytometry will be used to validate Ab specificity with human cancer cell lines. Complement-mediated cell killing (cytotoxicity) analysis using human tumor cell lines, will reveal Ab function. Finally, assessing and evaluating the efficacy of select TACA-PS A1 constructs to treat cancer will be demonstrated through in vivo experiments utilizing murine models grafted with the TA3/Ha murine breast carcinoma. The broader impact of the proposed research aims to demonstrate how entirely carbohydrate-based vaccines elicit cellular and humoral responses in the immune system. This research program will contribute to improved methodologies for the field of glycoconjugation chemistry, a better understanding of the role of PS A1 as an immunogen in eliciting immune responses, and assessment and evaluation of cancer specific constructs that are rationally designed. This project will, therefore, have an important impact on cancer treatment by providing a novel carbohydrate-based agent for preventive and therapeutic medicine and increase our understanding of glycoimmunology. We also expect this program to be broadly applicable to other areas of vaccine research.

描述（由申请人提供）：该研究计划的主要目标是开发实用的合成方法，用于修饰独特的荚膜两性离子多糖（ZPS）、PS A1，并缀合肿瘤相关碳水化合物抗原（TACA）以开发完全碳水化合物疫苗构建体。 PS A1 是一种结构明确的多糖，含有从厌氧菌脆弱拟杆菌 NCTC 9343 的荚膜多糖复合物 (CPC) 中分离出来的重复四糖核心。已确定，PS A1 可以通过诱导免疫系统激发细胞和体液免疫功能。 T 细胞和 B 细胞的参与影响了辅助反应和记忆反应事件——这是一个范式转变的发现。目前尚不完全清楚的是，完全基于碳水化合物的构建体作为肿瘤疫苗开发的糖蛋白缀合物的替代方法的免疫原性。因此，本研究计划旨在通过三个具体目标来研究和评估创新疫苗构建体：1) 开发有效的缀合方法，用于构建结构明确的 TACA-PS A1 缀合疫苗，2) 研究TACA-PS A1 在小鼠中缀合物，并评估 TACA-PS A1 产生的抗体对人类癌细胞系的特异性和细胞毒性，以及 3) 在体内评估TACA-PS A1 激活免疫来治疗肿瘤。从小鼠免疫接种中获得的抗血清将用于通过 ELISA 测定总抗体 (Ab) 滴度和 Ab 同种型，从而传递有关免疫的信息。此外，ELISA 将用于确定针对缀合 TACA 的抗体特异性。流式细胞术将用于验证抗体对人类癌细胞系的特异性。使用人类肿瘤细胞系进行补体介导的细胞杀伤（细胞毒性）分析将揭示 Ab 功能。最后，将通过利用移植有 TA3/Ha 小鼠乳腺癌的小鼠模型的体内实验来证明评估和评价所选 TACA-PS A1 构建体治疗癌症的功效。拟议研究的更广泛影响旨在证明完全基于碳水化合物的疫苗如何引发免疫系统中的细胞和体液反应。该研究计划将有助于改进糖缀合化学领域的方法，更好地了解 PS A1 作为免疫原在引发免疫反应中的作用，以及对合理设计的癌症特异性结构进行评估和评价。因此，该项目将为预防和治疗医学提供一种新型的基于碳水化合物的药物，并增加我们对糖免疫学的理解，从而对癌症治疗产生重要影响。我们还期望该计划能够广泛适用于疫苗研究的其他领域。